MYB2 Is Important for Tapetal PCD and Pollen Development by Directly Activating Protease Expression in Arabidopsis.

Tapetal programmed cell death (PCD) is a complex biological process that plays an important role in pollen formation and reproduction. Here, we identified the MYB2 transcription factor expressed in the tapetum from stage 5 to stage 11 that was essential for tapetal PCD and pollen development in Arabidopsis thaliana. Downregulation of MYB2 retarded tapetal degeneration, produced defective pollen, and decreased pollen vitality. EMSA and transcriptional activation analysis revealed that MYB2 acted as an upstream activator and directly regulated expression of the proteases CEP1 and ßVPE. The expression of these proteases was lower in the buds of the myb2 mutant. Overexpression of either/both CEP1 or/and ßVPE proteases partially recover pollen vitality in the myb2 background. Taken together, our results revealed that MYB2 regulates tapetal PCD and pollen development by directly activating expression of the proteases CEP1 and ßVPE. Thus, a transcription factor/proteases regulatory and activated cascade was established for tapetal PCD during another development in Arabidopsis thaliana. Highlight: MYB2 is involved in tapetal PCD and pollen development by directly regulating expression of the protease CEP1 and ßVPE and establishes a transcription factor/proteases regulatory and activated cascade.

Quantitative proteomics analysis of tomato growth inhibition by ammonium nitrogen.

As a single nitrogen source, ammonium (NH4+) can inhibit the growth of plants, especially when applied in excess. Tandem mass tag (TMT) quantitative proteomics technology was employed in the current study to explore and analyze the mechanisms of ammonium-induced inhibition. F1 tomato (Lycopersicon esculentum Mill) was used in this study. Seedlings at the four leaf-stages grown in a greenhouse were irrigated using nutrient solution with NH4+-N as single nitrogen source (15 mmol L-1, single NO3--N as control) for 5 weeks. Compared to the control, the root biomass of NH4+-N-treated seedlings decreased by 50%. In addition, NH4+ content in roots was 2.83-fold increased and soluble sugar and protein contents were increased. However, the starch content did not change significantly. The activities of glutamine synthetase (GS), glutamate synthetase (GOGAT) and glutamate dehydrogenase (GDH), which are involved in ammonium assimilation, were increased, and glutamine (Gln) content was also increased. However, glutamate (Glu) content, which is important for amino transfer, did not significantly increase. Ammonium assimilation was inhibited. Root quantitative proteomics showed that carbonic anhydrase Q5NE21 was significantly downregulated. Although K4BPV5 and K4D9J3 proteins, which improve ammonium assimilation, were upregulated, ammonium assimilation was limited. In addition, NH4+ accumulated, which is likely due to Q5NE21 downregulation. Meanwhile, cell wall metabolism related to phenylpropanoid biosynthesis was altered due to the accumulation of NH4+ levels. Subsequently, tomato root growth was inhibited.

The cysteine protease CEP1, a key executor involved in tapetal programmed cell death, regulates pollen development in Arabidopsis.

Tapetal programmed cell death (PCD) is a prerequisite for pollen grain development in angiosperms, and cysteine proteases are the most ubiquitous hydrolases involved in plant PCD. We identified a papain-like cysteine protease, CEP1, which is involved in tapetal PCD and pollen development in Arabidopsis thaliana. CEP1 is expressed specifically in the tapetum from stages 5 to 11 of anther development. The CEP1 protein first appears as a proenzyme in precursor protease vesicles and is then transported to the vacuole and transformed into the mature enzyme before rupture of the vacuole. cep1 mutants exhibited aborted tapetal PCD and decreased pollen fertility with abnormal pollen exine. A transcriptomic analysis revealed that 872 genes showed significantly altered expression in the cep1 mutants, and most of them are important for tapetal cell wall organization, tapetal secretory structure formation, and pollen development. CEP1 overexpression caused premature tapetal PCD and pollen infertility. ELISA and quantitative RT-PCR analyses confirmed that the CEP1 expression level showed a strong relationship to the degree of tapetal PCD and pollen fertility. Our results reveal that CEP1 is a crucial executor during tapetal PCD and that proper CEP1 expression is necessary for timely degeneration of tapetal cells and functional pollen formation.

Sex- and season-dependent differences in telomere length and telomerase activity in the leaves of ash and willow.

Telomeres and telomerase have important biological functions and can protect chromosome ends. In this study, sex- and season-dependent changes in telomere length and telomerase activity in ash and willow were analyzed. A statistical analysis showed that the telomere lengths of male and female trees differed significantly (P < 0.05). In ash, the telomere lengths of female trees were shorter than those of male trees. In willow, the telomere lengths of female trees were longer than those of male trees. During the annual developmental cycle, the telomere lengths of male and female ash and willow increased from April to May (P < 0.05), remained stable from May to August (P > 0.05), and decreased significantly in September and October (P < 0.05). Additionally, telomerase activities could be detected in both male and female ash and willow trees from April to October. Our results show that the telomere lengths changed according to season and sex in ash and willow. Telomere length did not have a direct positive correlation with telomerase activity.